Evaluation On Main Biological Characters Of Host-plant Haloxylon Ammodendron And Haloxylon Persicum Of Cistanche Deserticol

Abstract: Haloxylon ammodendron and Haloxylon persicum, belonging to the family Amaranthaceae, genus Haloxylon Bunge, serve as host plants for Cistanche deserticola. They are widely distributed in the arid zones of China. In this study, both species' biological characteristics and agronomic traits were mainly compared by field observation and molecular identification. The results showed significant differences in phenotypic characters, pollen structure, and agronomic traits. Notably, the fruit wing length and assimilation branch length in H. ammodendron were significantly lower than those of H. persicum. The pollen volume and surface particle structure of H. ammo dendron were significantly smaller than those of H. persicum. Additionally, the pollen net of H. ammodendron exhibited a deep structure, while it was shallow and most of it protruded outwards in H. persicum pollens. The correlational analysis revealed significant positive correlations between the agronomic indexes of both H. ammodendron and H. persicum and climate factors such as temperature and humidity. Water and light are the primary limiting factors for the geographic distribution of H. persicum in the Junggar Basin of Xinjiang, while environmental adaptations of H. ammodendron were found to be stronger. H. ammodendron and H. persicum can be effectively identified by ITS barcode, and both of them can be parasitized by C. deserticola. However, the survival rate of C. deserticola was different. According to their biological characteristics, cultivating H. ammodendron is with priority in the production of cistanches herba. This study provided a scientific basis for the identification and conservation of Haloxylon, as well as the selection of optimal hosts for C. deserticola. 

Keywords: Haloxylon ammodendron;Haloxylon persicum;Cistanche deserticola;Pollen;ITS

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Plants of the genus Haloxylon ammodendron (Haloxylon Bunge) are mainly distributed in regions from the Mediterranean to Central Asia worldwide. In my country, there is Haloxylon ammodendron [Haloxylonammdendron (C. A. Mey.) Bunge] and Haloxylon persicum Bunge ex Boiss. et Buhse 2 species[1]. Among them Haloxylon ammo dendron

It is widely distributed in Xinjiang, Qinghai, Gansu, Inner Mongolia Ningxia, and other regions in my country. It is a common tree species in desert areas and mainly grows in gravel deserts, clay deserts, fixed and semi-fixed sandy lands, and saline-alkali lands; while Haloxylon ammo dendron is mainly distributed in my country In the western region of the Junggar Basin in Xinjiang, most of the

In mobile, semi-fixed, and fixed dune areas [2]. Due to its drought resistance and strong adaptability, Haloxylon ammodendron grows well in desert areas with poor soil in northwest my country and plays an important role in windbreak and sand fixation and maintenance of ecological balance [3]. In addition, Cistanche deserticola, which is parasitic on the roots of Haloxylon ammo dendron

(Cistanche deserticola Y. C. Ma) is a famous tonic Chinese medicinal material in my country [4]. In recent years, large-scale predatory mining has brought wild Cistanche deserticola resources to the verge of extinction and seriously damaged the ecological environment. Today, Cistanche deserticola and its hosts Haloxylon ammodendron and Haloxylon ammodendron alba have been included in the national secondary protected plant list [5]. With the rise of the international "return to nature" trend, and the fact that Cistanche deserticola has recently been officially included in the catalog of medicinal and food homologous products by the National Health Commission and the State Administration for Market Regulation, the market demand for Cistanche deserticola is constantly increasing. At this stage, Cistanche deserticola is being evaluated to host Haloxylon ammonium germplasm resources and artificial planting research is of great significance.

Haloxylon ammodendron plants in my country have a wide geographical distribution and rich intraspecific genetic diversity. Screening of excellent germplasm that grows quickly and has a high parasitism rate of Cistanche deserticola can effectively improve the yield and quality of Cistanche deserticola. At present, there are many studies on the cultivation and physiological ecology of Haloxylon ammodendron and Haloxylon ammodendron and Haloxylon ammodendron [6-7], but there are relatively few studies on the comparison of biological traits of Haloxylon ammodendron and Haloxylon ammodendron and Haloxylon ammodendron and the differences in parasitism of Cistanche deserticola between the two. Chen Yuchao et al. [8] used microslicing technology to study and showed that the root structure of the two is the same, consisting of the epidermis, cortex, and vascular column, and the proportion of cortical parenchyma cells in the root is larger. Research by Sun Fangfang et al. [9] showed that the genetic differentiation between Haloxylon ammodendron and Haloxylon ammodendron and Haloxylon ammodendron species is obvious, and the molecular system relationship is relatively clear. Based on the combination of ITS and cpDNA sequences, research on the phylogeny, genetics, and evolution of the two species can be carried out. To comprehensively evaluate the diversity of Haloxylon ammodendron and Haloxylon ammodendron albicans germplasm resources that host Cistanche deserticola, and screen the excellent host germplasm resources of Cistanche deserticola, this study carried out the Haloxylon ammodendron and Haloxylon ammodendron and Haloxylon ammodendron albicans germplasm resources collected in the early stage of the research group. Analysis of plant morphological characteristics, pollen characteristics, agronomic traits, and parasitism differences of Cistanche deserticola of Haloxylon ammodendron. The research results will provide a scientific basis for the identification, introduction, and selection of excellent host germplasm of Cistanche deserticola.

1 Materials and methods

1.1 Test materials

Haloxylon ammodendron and Haloxylon ammodendron seeds were collected from the Haloxylon ammodendron Forest National Nature Reserve in Ganjia Lake, Xinjiang. The collected seeds were dried and impurities removed and stored in a 4°C refrigerator for later use. The pollen of Haloxylon ammodendron and Haloxylon ammodendron and Haloxylon ammodendron were collected from the Herba Cistanches planting base in Yongning County, Ningxia. The flower buds of 10 Haloxylon ammodendron or Haloxylon ammodendron each were collected during the peak flowering period, and then naturally dried and stored in a -20°C refrigerator for later use. The Haloxylon ammodendron and Haloxylon ammodendron albicans cultivation test samples came from the Cistanche deserticola planting base in Yongning County, Ningxia, and the wild Cistanche deserticola host materials were collected from the Haloxylon ammodendron Forest National Nature Reserve in Ganjia Lake, Xinjiang. All experimental materials used were identified by researcher Chen Jun of the Institute of Medicinal Plants, Chinese Academy of Medical Sciences, and are stored in the germplasm resource bank of the Institute of Medicinal Plants, Chinese Academy of Medical Sciences.

1.2 Method

1.2.1 Comparison of biological characteristics

A comparison of plant morphology was carried out using observation and measurement methods. The seed characteristics were compared by randomly selecting 30 Haloxylon ammodendron or Haloxylon ammodendron seeds from different sampling points, and vernier calipers were used to measure the seed diameter, thickness, fruit wing length, and other indicators; the thousand-seed weight was selected using the hundred-seed method; the seeds were placed on a 2-layer paved surface. Add an appropriate amount of deionized water to a petri dish on filter paper, and culture it at 25°C. Based on the growth of the seeds, determine the investigation time for germination potential and germination rate.

The pollen characteristics were compared according to the method of Qiao Qi et al. [10]. The pollen was sprayed with gold using a JFC-1600 ion sputtering instrument. The morphological characteristics of the pollen were observed under a scanning electron microscope (JSM-6510LV). The size, polar axis length, and length of the pollen were measured. Equatorial axis width, mesh ridge width, mesh size, knob-like protrusions, and other indicators. The size of pollen is expressed by P╳E value, and the shape of pollen is expressed by P/E, where P/E>2 is super long spherical, and 1.14<P/E<2 is long spherical. At the same time, observe the pollen shape, germination pore type, and outer wall decoration. etc.[11].

1.2.2 Analysis of Agronomic Indicators

Agronomic character tests of Haloxylon ammodendron and Haloxylon ammodendron and Haloxylon ammodendron albicans were conducted at the Materia Medica Cistanches planting base in Yongning County, Ningxia. Before sowing, the test soil was deeply plowed, and 3000kg/666.7 m2 organic fertilizer was applied as the base fertilizer. The surface of the soil was raked and the seedbed was laid out. The length and width of the seedbed were 30 m╳3 m. Use 0.3% before sowing for Haloxylon ammodendron and Haloxylon ammodendron alba.

Soak in potassium permanganate solution for 30 minutes, rinse with clean water, mix well with 5 times the volume of fine sand, and sow in wide strips with a seeding rate of 5 kg/666.7m2. According to the method of Li Miao et al. [12], indicators including plant height, root length, number of primary branches, number of branches, stem base diameter, plant fresh weight, and plant dry weight were measured.

1.2.3 Parasitism rate of Cistanche deserticola in the field and identification of wild Cistanche deserticola host species

Haloxylon ammodendron and Haloxylon ammodendron were selected for the Cistanche deserticola inoculation and parasitism test. Except for the different host plants, the planting management was carried out according to the Cistanche deserticola cultivation technical regulations formulated by the relevant personnel of this study [13]. Mix Cistanche deserticola seeds with fine sand and spread them into a 50 cm deep inoculation trench. One year later, observe the parasitism of Cistanche deserticola inoculation, and calculate the number of Cistanche deserticola in each inoculation trench and the parasitism rate. The parasitism rate of Cistanche deserticola is the percentage of the number of parasitic Cistanche deserticola plants to the total number of plants in the experimental area. The field collection experiment was conducted in Ganjia Lake Haloxylon Ammodendron National Nature Reserve in Xinjiang. Experimental materials of Cistanche deserticola and its parasitic roots were collected from 28 sampling points. Based on the host identification method of halophytic Cistanche deserticola studied by Shen Liang et al. [14] ITS molecular identification of Cistanche deserticola parasitic roots

1.2.4 Data statistical analysis

Excel 2013 and SPSS 20.0 statistical software were used to conduct a single-factor analysis of variance on agronomic traits of Haloxylon ammodendron and Haloxylon ammodendron and Haloxylon ammodendron albus, pollen polar axis length (P), equatorial axis width (E), P╳E and P/E, mesh size and other indicators. and related analysis. MEGA5.0 was used to conduct a comparative analysis of Cistanche deserticola host bases, and a phylogenetic NJ (neighbor-joining) tree was constructed. The nucleotide substitution model was the Kimura-2-parameter model, and the bootstrap value (bootstrap) was 1000. Check the support rate of each branch. The samples were identified and analyzed based on the sequences of Haloxylon ammodendron and Haloxylon ammodendron in the GeneBank database. The GeneBank accession number of the Haloxylon ammodendron ITS reference sequence is HM131632.1, and the GeneBank accession number of the Haloxylon ammodendron ITS reference sequence is HM131633.1.